The final week of February, 2019 (Feb. 25 - Mar 1), iLab Operations Software will be conducting an internal event and will be staffed at significantly reduced levels. iLab Support will prioritize critical issues first (i.e. issues that prevent key workflows from completing) and to escalate an issue, please add "Urgent" to the ticket/ email subject or press "1", when prompted, on the iLab Support phone (regional numbers can be found here). Regular support tickets will be monitored periodically and processed in order of submission starting March 4th, 2019. During this time, you can also refer to the iLab Help Site and iLab Status Page for site information. Please note that this will not affect standard Core Facility Operations.

Stable Isotope and Metabolomics Core

Overview

The Stable Isotope and Metabolomics Core can provide targeted and untargeted metabolite profiling of plasma or tissue, and the rate of change of substrates (flux), using stable isotopes and mass spectrometry. The core has a variety of in vitro and in vivo metabolic methodologies that can determine substrate flux dynamics and metabolite profiles at the organelle, cellular, tissue and whole body levels. In collaboration with other Diabetes Research Center cores, the application of mass spectrometric and other methodologies allows the elucidation of metabolic mechanisms underlying disorders in fuel homeostasis. The metabolic role of candidate molecules can be specifically delineated in vivo and in vitro, using a step-by-step hypothesis-driven approach in animals, cell lines and other models.

The pump contains 50 mg of [U-13C6] glucose, each dissolved in 200 uL of water. The mini-pump is inserted in the subcutaneous space, and therefore infusion conditions approximate those of the venous-arterial (V-A) mode of infusion and sampling. Mass isotopomer distribution analysis is used for calculation of glucose production and recycling.

Bile acids are synthesized from cholesterol through both classical and alternative pathways. In the alternative pathway, the side chain oxidation of cholesterol precedes the steroid ring modifications, first yielding 24-, 25-, and 27-hydroxycholesterol metabolites, opposite to the process in the classical pathway. The alternative and classical pathway bile acids share the primary bile acid chenodeoxycholic acid, with 12-a-hydroxylation of chenodeoxycholic acid via CYP8B1to cholic acid. Modifications of bile acids can affect their properties and their ability to activate bile acid receptors. Dysregulation of bile acid synthesis can be seen in inborn errors of metabolism, insulin resistance, hepatocellular Ca and chronic ethanol consumption. Perturbations in the microbiome also affect bile acid pool size and composition (see references below). This panel surveys conditions of bile acid dysregulation. Please see our website for more information about the pertinent metabolic pathways for this module.

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Module # 6 - Acyl-CoA analysis

This module includes acetyl CoA, malonyl CoA, and succinyl CoA, in order to interface with acyl carnitine analysis in Module #1, and TCA cycle measurements in Modules # 3 and 7.

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Module # 7 - GC/MS Small Metabolite Screen

GC/MS will identify about 80 to 120 small metabolites based on the difference of samples, in the combined glycolytic/gluconeogenic, pentose and TCA cycle pathways. For an example by the SIMC facility, see:

The targeted metabolomics approach in this assay is based on measurements with the AbsoluteIDQ p180 kit (BIOCRATES Life Sciences AG, Innsbruck, Austria). This method allows simultaneous quantification of 188 metabolites in plasma, tissues or cell pellets using liquid chromatography and flow injection analysis–mass spectrometry. For an example of this module’s utility, see: Wang-Sattler et al Novel biomarkers for pre-diabetes identified by metabolomics, Molecular Systems Biology 8; 615; doi:10.1038/msb.2012.43. Please see our website for more information about the pertinent metabolic pathways for this module.

Inquire

Module #2 - Choline, Betaine, TMAO and Creatinine

Choline and betaine are essential nutrients. Choline is important for brain function, liver health, reproduction, and fetal and infant development. Betaine acts as a methyl donor in the liver and as an important osmolyte to protect the cells of medulla in the kidney. Their gut microbial metabolite trimethylamine N-oxide (TMAO) was considered non-toxic, but it recently has been associated with increased risk for cardiovascular disease. This finding has stimulated a growing interest in analyzing these compounds in bio fluids and studying their associations with human health and diseases. This assay allows simultaneous quantification of free choline, betaine, TMAO and creatinine in plasma, urine or tissue samples. The analysis is performed using liquid chromatography-stable isotope dilution-multiple reaction monitoring mass spectrometry (LC-SID-MRM/MS). Please see our website for more information about the pertinent metabolic pathways for this module.

Inquire

Sample preparation - Tissue sample powder and weight

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Setup Fee - Weekdays after business hours

Covers staff overtime for cleaning and setup after business hours.

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Setup Fee - Weekdays during business hours (Waived for DRC Members)

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Setup Fee - Weekends

Covers staff overtime for cleaning and setup for weekend use.

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Training Fee

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XF24 - 3 Extracellular Flux Analysis Plate & Reagent Fee

Note: This is only the price for the plate and reagents. Seahorse services are priced at an hourly rate plus the cost of the plate and reagents.

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XF96 Extracellular Flux Analysis Plate & Reagent Fee

Note: This is only the price for the plate and reagents. Seahorse services are priced at an hourly rate plus the cost of the plate and reagents.

The targeted metabolomics approach in this assay is based on measurements with the AbsoluteIDQ p180 kit (BIOCRATES Life Sciences AG, Innsbruck, Austria). This method allows simultaneous quantification of 188 metabolites in plasma, tissues or cell pellets using liquid chromatography and flow injection analysis–mass spectrometry. For an example of this module’s utility, see: Wang-Sattler et al Novel biomarkers for pre-diabetes identified by metabolomics, Molecular Systems Biology 8; 615; doi:10.1038/msb.2012.43. Please see our website for more information about the pertinent metabolic pathways for this module.

Inquire

Module #2 - Choline, Betaine, TMAO and Creatinine

Choline and betaine are essential nutrients. Choline is important for brain function, liver health, reproduction, and fetal and infant development. Betaine acts as a methyl donor in the liver and as an important osmolyte to protect the cells of medulla in the kidney. Their gut microbial metabolite trimethylamine N-oxide (TMAO) was considered non-toxic, but it recently has been associated with increased risk for cardiovascular disease. This finding has stimulated a growing interest in analyzing these compounds in bio fluids and studying their associations with human health and diseases. This assay allows simultaneous quantification of free choline, betaine, TMAO and creatinine in plasma, urine or tissue samples. The analysis is performed using liquid chromatography-stable isotope dilution-multiple reaction monitoring mass spectrometry (LC-SID-MRM/MS). Please see our website for more information about the pertinent metabolic pathways for this module.

Bile acids are synthesized from cholesterol through both classical and alternative pathways. In the alternative pathway, the side chain oxidation of cholesterol precedes the steroid ring modifications, first yielding 24-, 25-, and 27-hydroxycholesterol metabolites, opposite to the process in the classical pathway. The alternative and classical pathway bile acids share the primary bile acid chenodeoxycholic acid, with 12-a-hydroxylation of chenodeoxycholic acid via CYP8B1to cholic acid. Modifications of bile acids can affect their properties and their ability to activate bile acid receptors. Dysregulation of bile acid synthesis can be seen in inborn errors of metabolism, insulin resistance, hepatocellular Ca and chronic ethanol consumption. Perturbations in the microbiome also affect bile acid pool size and composition (see references below). This panel surveys conditions of bile acid dysregulation. Please see our website for more information about the pertinent metabolic pathways for this module.

Inquire

Module # 6 - Acyl-CoA analysis

This module includes acetyl CoA, malonyl CoA, and succinyl CoA, in order to interface with acyl carnitine analysis in Module #1, and TCA cycle measurements in Modules # 3 and 7.

Inquire

Module # 7 - GC/MS Small Metabolite Screen

GC/MS will identify about 80 to 120 small metabolites based on the difference of samples, in the combined glycolytic/gluconeogenic, pentose and TCA cycle pathways. For an example by the SIMC facility, see:

The pump contains 50 mg of [U-13C6] glucose, each dissolved in 200 uL of water. The mini-pump is inserted in the subcutaneous space, and therefore infusion conditions approximate those of the venous-arterial (V-A) mode of infusion and sampling. Mass isotopomer distribution analysis is used for calculation of glucose production and recycling.